Title The Emerging Role of Rhinoviruses in Lower Respiratory Tract Infections in Children - Clinical and Molecular Epidemiological Study From Croatia, 2017-2019.
Author Ljubin-Sternak, Suncanica; Mestrovic, Tomislav; Ivkovic-Jurekovic, Irena; Kolaric, Branko; Slovic, Anamarija; Forcic, Dubravko; Tot, Tatjana; Mijac, Maja; Vranes, Jasmina
Journal Front Microbiol Publication Year/Month 2019
PMID 31849887 PMCID PMC6901631
Affiliation + expend 1.Molecular Microbiology Department, Dr. Andrija Stampar Teaching Institute of Public Health, Zagreb, Croatia.

Rhinoviruses (RVs) are increasingly implicated not only in mild upper respiratory tract infections, but also in more severe lower respiratory tract infections; however, little is known about species diversity and viral epidemiology of RVs among the infected children. Therefore, we investigated the rhinovirus (RV) infection prevalence over a 2-year period, compared it with prevalence patterns of other common respiratory viruses, and explored clinical and molecular epidemiology of RV infections among 590 children hospitalized with acute respiratory infection in north-western and central parts of Croatia. For respiratory virus detection, nasopharyngeal and pharyngeal flocked swabs were taken from each patient and subsequently analyzed with multiplex RT-PCR. To determine the RV species in a subset of positive children, 5\'UTR in RV-positive samples has been sequenced. Nucleotide sequences of referent RV strains were retrieved by searching the database with Basic Local Alignment Tool, and used to construct alignments and phylogenetic trees using MAFFT multiple sequence alignment tool and the maximum likelihood method, respectively. In our study population RV was the most frequently detected virus, diagnosed in 197 patients (33.4%), of which 60.4% was detected as a monoinfection. Median age of RV-infected children was 2.25 years, and more than half of children infected with RV (55.8%) presented with lower respiratory tract infections. Most RV cases were detected from September to December, and all three species co-circulated during the analyzed period (2017-2019). Sequence analysis based on 5\'UTR region yielded 69 distinct strains; the most prevalent was RV-C (47.4%) followed by RV-A (44.7%) and RV-B (7.9%). Most of RV-A sequences formed a distinct phylogenetic group; only strains RI/HR409-18 (along with a reference strain MF978777) clustered with RV-C strains. Strains belonging to the group C were the most diverse (41.6% identity among strains), while group B was the most conserved (71.5% identity among strains). Despite such differences in strain groups (hitherto undescribed in Croatia), clinical presentation of infected children was rather similar. Our results are consistent with newer studies that investigated the etiology of acute respiratory infections, especially those focused on children with lower respiratory tract infections, where RVs should always be considered as potentially serious pathogens.

StrainID RV Species Serotype Length(nt) Country Year Strain Name
ANC02599 A None 335 Croatia 2017 RI/HR133-17
ANC02604 A None 332 Croatia 2017 RI/HR118-17
ANC02609 A None 331 Croatia 2018 RI/HR554-18
ANC02614 A None 331 Croatia 2017 RI/HR60-17
ANC02619 A None 331 Croatia 2018 RI/HR483-18
ANC02624 A None 332 Croatia 2019 RI/HR646-19
ANM02600 A None 332 Croatia 2018 RI/HR409-18
ANM02605 A None 332 Croatia 2017 RI/HR197-17
ANM02610 A None 332 Croatia 2018 RI/HR584-18
ANM02615 A None 334 Croatia 2018 RI/HR294-18
ANM02620 A None 332 Croatia 2017 RI/HR186-17
ANM02625 A None 332 Croatia 2018 RI/HR268-18
ANP02597 A None 335 Croatia 2018 RI/HR507-18
ANP02602 A None 334 Croatia 2018 RI/HR553-18
ANP02607 A None 332 Croatia 2018 RI/HR561-18
ANP02612 A None 331 Croatia 2018 RI/HR601-18
ANP02617 A None 333 Croatia 2017 RI/HR104-17
ANP02622 A None 331 Croatia 2017 RI/HR91-17
ANS02598 A None 334 Croatia 2018 RI/HR430-18
ANS02603 A None 332 Croatia 2018 RI/HR524-18
ANS02608 A None 332 Croatia 2019 RI/HR791-19
ANS02613 A None 331 Croatia 2017 RI/HR70-17
ANS02618 A None 333 Croatia 2017 RI/HR126-17
ANS02623 A None 332 Croatia 2019 RI/HR848-19
ANZ02601 A None 334 Croatia 2018 RI/HR518-18
ANZ02606 A None 333 Croatia 2018 RI/HR602-18
ANZ02611 A None 331 Croatia 2017 RI/HR58-17
ANZ02616 A None 332 Croatia 2018 RI/HR501-18
ANZ02621 A None 331 Croatia 2017 RI/HR77-17
BNC00424 B None 329 Croatia 2018 RI/HR520-18
BNM00425 B None 335 Croatia 2017 RI/HR218-17
BNP00422 B None 333 Croatia 2019 RI/HR668-19
BNS00423 B None 331 Croatia 2018 RI/HR490-18
BNZ00426 B None 335 Croatia 2017 RI/HR105-17
CNC01329 C None 338 Croatia 2018 RI/HR559-18
CNC01334 C None 337 Croatia 2017 RI/HR127-17
CNC01339 C None 333 Croatia 2017 RI/HR90-17
CNC01344 C None 333 Croatia 2018 RI/HR577-18
CNC01349 C None 305 Croatia 2017 RI/HR164-17
CNC01354 C None 335 Croatia 2018 RI/HR592-18
CNC01359 C None 331 Croatia 2018 RI/HR568-18
CNM01325 C None 338 Croatia 2019 RI/HR606-19
CNM01330 C None 332 Croatia 2017 RI/HR214-17
CNM01335 C None 306 Croatia 2017 RI/HR160-17
CNM01340 C None 333 Croatia 2017 RI/HR137-17
CNM01345 C None 333 Croatia 2017 RI/HR132-17
CNM01350 C None 333 Croatia 2017 RI/HR103-17
CNM01355 C None 332 Croatia 2017 RI/HR148-17
CNP01327 C None 337 Croatia 2017 RI/HR204-17
CNP01332 C None 311 Croatia 2017 RI/HR119-17
CNP01337 C None 333 Croatia 2019 RI/HR617-19
CNP01342 C None 333 Croatia 2017 RI/HR79-17
CNP01347 C None 332 Croatia 2017 RI/HR75-17
CNP01352 C None 335 Croatia 2017 RI/HR52-17
CNP01357 C None 331 Croatia 2019 RI/HR839-19
CNS01328 C None 338 Croatia 2019 RI/HR649-19
CNS01333 C None 336 Croatia 2017 RI/HR76-17
CNS01338 C None 332 Croatia 2017 RI/HR71-17
CNS01343 C None 333 Croatia 2019 RI/HR751-19
CNS01348 C None 335 Croatia 2018 RI/HR488-18
CNS01353 C None 332 Croatia 2017 RI/HR74-17
CNS01358 C None 333 Croatia 2017 RI/HR205-17
CNZ01326 C None 334 Croatia 2017 RI/HR210-17
CNZ01331 C None 335 Croatia 2018 RI/HR496-18
CNZ01336 C None 333 Croatia 2019 RI/HR626-19
CNZ01341 C None 332 Croatia 2017 RI/HR97-17
CNZ01346 C None 332 Croatia 2019 RI/HR664-19
CNZ01351 C None 335 Croatia 2018 RI/HR509-18
CNZ01356 C None 331 Croatia 2017 RI/HR115-17
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